A peptide vaccine administered transcutaneously together with cholera toxin elicits potent neutralising anti-FMDV antibody responses

In this study a synthetic peptide representing residues 141-159 from the GH loop of VP1 protein of foot-and-mouth disease virus was tested for its capacity to elicit virus neutralising antibodies in mice after transcutaneous immunisation. Topical application of the peptide conjugated to bovine serum albumin together with cholera toxin as an adjuvant elicited anti-peptide antibody responses with strong virus neutralising activity. The combination of cholera toxin with an immunostimulatory CpG motif resulted in the induction of IgG1 and IgG2a anti-peptide antibodies with significantly enhanced virus neutralising activity. To shed more light on the mechanisms of cholera toxin adjuvanticity we demonstrated its binding to keratinocytes via GM(1)-gangliosides. This was followed by an increase of the intracellular cAMP and the rapid diffusion of cholera toxin throughout the epidermis. These findings demonstrate that peptide-based vaccines when combined with the appropriate adjuvant(s) can elicit potent virus neutralising antibody responses after transcutaneous immunisation. However, experiments in target species will be required to confirm the potential of this simple vaccination procedure in livestock.     

Lapine rotaviruses of the genotype P[22] are widespread in Italian rabbitries

An epidemiological survey was carried out to investigate the distribution of the VP7 and VP4 specificities of lapine rotaviruses (LRVs) in rabbitries from different geographical regions of Italy. Almost all the strains were characterized as P[22],G3, confirming the presence of the newly-recognized rotavirus P[22] VP4 allele in Italian rabbits. Only one P[14],G3 LRV strain was identified and two samples contained a mixed (P[14] + [22],G3) rotavirus infection. All the LRV strains analyzed exhibited a genogroup I VP6 specificity and a long dsRNA electropherotype. However, one of the P[14],G3 strains possessed a super-short pattern. Altogether, these data highlight the epidemiological relevance of the P[22] LRVs in Italian rabbitries.     

Virological and molecular characterization of a mammalian orthoreovirus type 3 strain isolated from a dog in Italy

A mammalian orthoreovirus (MRV) strain was isolated from a pup with fatal diarrhea, which had a concurrent infection by canine parvovirus type 2. The reovirus isolate showed an atypical hemagglutination pattern and a retarded electrophoretic mobility of the S1 segment, which is characteristic of MRV type 3 (MRV-3). Assignment of the isolated virus to MRV-3 was confirmed by type-specific RT-PCR assays, targeting the S1 gene, and by subsequent sequence analysis of the PCR product. By phylogeny based on the S1 gene of several MRVs, the isolate fell into lineage E, along with the murine strain T3C9/61 and the bovine strains T3C18/61 and T3C31/59. Conversely, L1 sequences were found to segregate regardless of the viral type. A total of 110 fecal samples, 56 nasal and 31 ocular swabs from dogs with diarrhea or nasal/ocular discharge were tested by a nested-PCR assay specific for reoviruses, and no sample was found to contain MRV RNA, a finding that is apparently in contrast with the seroprevalence (25.77%) observed in dogs.     

Carmustine, vincristine, and prednisone in the treatment of canine lymphosarcoma

A chemotherapeutic protocol using carmustine in combination with vincristine and prednisone was tested in dogs with multicentric malignant lymphosarcoma. Of seven dogs treated, six (85.7%) achieved complete remission. A partial response occurred in one dog. Median survival time was 224 days (mean 386 days), and median duration of remission was 183 days (mean 323 days). Marked neutropenia was observed following carmustine administration. There were no significant alterations in platelets and red blood cell counts during treatment, and no abnormalities attributable to the chemotherapy were found in serum biochemical profiles. Results of this study showed that carmustine is an effective alternative option in the treatment of canine lymphosarcoma.     

SPINAL SUB ARACHNOID CYSTS IN 13 DOGS

Thirteen dogs, including 6 Rottweiler dogs, exhibiting clinical signs of spinal cord dysfunction and myelographically confirmed subarachnoid space enlargement were investigated. To characterize the lesions and to get a better understanding of their pathogenesis, different imaging techniques were used in association with explorative surgical procedures (12 dogs) and histopathologic techniques (5 dogs). All subjects underwent preoperative myelography, five of which were examined by computed tomography (CT) scanning and one by magnetic resonance imaging (MRI) as well as cerebrospinal fluid (CSF) flow measurement (velocimetry). Most animals were <12 months old (7/13 dogs) and Rottweilers were over-represented (6/13 dogs). The lesions were mainly located dorsally with respect to the spinal cord (10/13 dogs) and in the cranial cervical area (8/13 dogs). MRI suggested spinal cord deviation with signs of ventral leptomeningeal adhesion opposite the enlarged space. In one dog, velocimetry confirmed that the "cyst" was freely communicating with the surrounding CSF space. Surgical investigation confirmed leptomeninges-induced ventral adhesion in 4/5 dogs. Follow-up studies, carried out from 6 months to 2.5 years postoperatively, showed there was full recovery in 8/13 dogs. This study suggests that the compression of the spinal cord is possibly not caused by a cyst. Adhesion resulting from a combination of microtrauma and chronic inflammatory processes induces a secondary enlargement of the subarachnoid space and may be a significant causative factor in spinal cord compression and dysfunction. The over-representation of Rottweilers and the young age of the animals in the study suggest a possible genetic predisposition and an inherited etiology.     

Short-term evaluation of dorsal acetabular augmentation in 10 canine total hip replacements

OBJECTIVE: To report the use of dorsal acetabular augmentation (DAA) in canine total hip arthroplasty (THA) and to evaluate the clinical and radiographic outcome after a minimum of 1 year follow-up. STUDY DESIGN: Clinical study. ANIMALS: Nine dogs that had THA. METHODS: Ten hips requiring acetabular augmentation for optimum acetabular cup implantation for THA were evaluated retrospectively. The excised femoral head and neck was used as the corticocancellous bone graft for index THA surgeries, and the dorsal crest of the ipsilateral ilial wing was used as the bone graft in 1 dog that had a revision surgery. Clinical and radiographic follow-up examinations were performed at approximately 8 weeks and 1 year postoperatively. Long-term radiographic examinations included use of a standing dorsal acetabular rim (DAR) projection for assessment of graft-recipient boundary. Functional clinical outcome was assessed using direct patient evaluation and telephone interview. RESULTS: One dog failed to regain limb function after surgery. Suspected aseptic loosening of the acetabular cup was observed on radiographs approximately 12 weeks postoperatively. Excluding this single explantation, follow-up physical and radiographic examinations indicated good implant and graft positioning and normal function for all dogs. CONCLUSIONS: Acetabular augmentation permitted implantation of an appropriately sized acetabular cup while maintaining complete DAR coverage of the prosthesis. Nine out of 10 hips had a successful outcome with minimal radiographic and no functional abnormalities. Further follow-up is necessary to determine the long-term outcome of DAR augmentation in canine THA. CLINICAL RELEVANCE: Acetabular augmentation with an autogenous corticocancellous bone graft constructed from the femoral head and neck can provide complete DAR coverage to permit use of an appropriately sized acetabular cup in THA.     

Lactoferrin against Staphylococcus aureus Mastitis. Lactoferrin alone or in combination with penicillin G on bovine polymorphonuclear function and mammary epithelial cells colonisation by Staphylococcus aureus

Antibiotics should combine good antibacterial activity and the capacity to work in association with the host defence system. In this study, we have investigated the effects of bovine lactoferrin alone or in combination with penicillin G on the phagocytic activity of bovine polymorphonuclear leukocytes against Staphylococcus aureus. We have shown that susceptibility of S. aureus to phagocytosis was decreased in the presence of penicillin in the medium. In a kinetic study, lactoferrin alone did not affect phagocytosis but, when used with penicillin, it reversed the negative effect of this antibiotic on phagocytosis. In addition, in an epithelial invasion assay, lactoferrin alone or in combination with penicillin reduced the invasion of mammary epithelial cells in culture by S. aureus. Lactating female CD-1 mice were infected by intra-mammary delivery of a virulent penicillin-susceptible S. aureus strain and were then randomly assigned to treatments according to a 2 x 2 factorial design. In this mouse mastitis model, 2 days of systemic treatments with lactoferrin and/or penicillin did not lead to a total clearance of infection by S. aureus, but bacterial number was significantly reduced by treatments with lactoferrin or penicillin. These data suggest that bovine lactoferrin, alone or in combination with penicillin G, enhances S. aureus susceptibility to immuno-defense mechanisms, which can be beneficial in the treatment of S. aureus infections.     

Use of the impulse oscillometry system for testing pulmonary function during methacholine bronchoprovocation in horses

OBJECTIVE: To compare sensitivity of the impulse oscillometry system (IOS) with that of the conventional reference technique (CRT; ie, esophageal balloon method) for pulmonary function testing in horses. ANIMALS: 10 horses (4 healthy; 6 with recurrent airway obstruction [heaves] in remission). PROCEDURE: Healthy horses (group-A horses) and heaves-affected horses (group-B horses) were housed in a controlled environment. At each step of a methacholine bronchoprovocation test, threshold concentration (TC(2SD); results in a 2-fold increase in SD of a value) and sensitivity index (SI) were determined for respiratory tract system resistance (R(rs)) and respiratory tract system reactance (X(rs)) at 5 to 20 Hz by use of IOS and for total pulmonary resistance (RL) and dynamic lung compliance (C(dyn)), by use of CRT. RESULTS: Bronchoconstriction resulted in an increase in R(rs) at 5 Hz (R(5Hz)) and a decrease in X(rs) at all frequencies. Most sensitive parameters were X(rs) at 5 Hz (X(5Hz)), R(5Hz), and R(5Hz):R(10Hz) ratio; RL and the provocation concentration of methacholine resulting in a 35% decrease in dynamic compliance (PC(35)C(dyn)) were significantly less sensitive than these IOS parameters. The TC(2SD) for X(rs) at 5 and 10 Hz was significantly lower in group-B horses, compared with group-A horses. The lowest TC(2SD) was obtained for X(5Hz) in group-B horses and R(5Hz) in group-A horses. CONCLUSIONS AND CLINICAL RELEVANCE: In contrast to CRT parameters, IOS parameters were significantly more sensitive for testing pulmonary function.The IOS provides a practical and noninvasive pulmonary function test that may be useful in assessing subclinical changes in horses.     

Experimental model of Border Disease Virus infection in lambs: comparative pathogenicity of pestiviruses isolated in France and Tunisia

Pestiviruses have been isolated from live sheep pox Tunisian vaccines. Vaccination with these vaccines caused outbreaks of Border Disease in Tunisia. In order to study more precisely the pathogenicity of these isolates, three groups of eight four month old lambs from a pestivirus-free flock were infected by the intratracheal route with a French strain (AV) and two Tunisian isolates (SN3G and Lot21). Clinical, hematological, immunological and virological parameters were evaluated. The three groups developed mild fever and leucopaenia by day 3 to 6 post infection (pi). The differences in the weight curves were not significant. Viruses were isolated from the peripheral blood buffy coat cells by day 4 to 9 pi. Antibodies were present on day 16 pi following infection by the French strain and on day 21 pi with the Tunisian isolates. The results demonstrated that SN3G and Lot21 are almost similar to the French strain used as the reference strain. In field conditions, they could induce economical losses in naive flocks, alone or in association with other pathogens.     

Lectinhistochemical and cytometrical evaluation of the corpus luteum of the rat at the end of pregnancy

Most studies on the biochemistry and structure of the corpus luteum have focused on elucidating the processes of progesterone synthesis and release. In the present work, the histochemical composition of the corpus luteum of the rat was evaluated using lectinhistochemistry on rats at the end of pregnancy (days 18-23). We also analysed the morphology of the luteal cells, to characterize the changes attributable to regression in this organ. Seven biotinylated lectins were used (CON-A, WGA, DBA, SBA, PNA, RCA and UEA-I) following pre-set protocols (ABC method). The average diameter and area of the cells and their nuclei were measured. High reactivity of the luteal cells was observed with CON-A and a lower reactivity with WGA. The capillary endothelium gave positive reactivity with WGA and to a certain extent with SBA, PNA and RCA. Vesicular structures were intensely stained with DBA, and were more abundant in sections from animals with more advanced pregnancy, which could be attributable to cellular debris, on the basis of their morphologic characteristics. There were no significant differences among the cytometric variables analysed in comparisons of the values corresponding to the different days of gestation. These observations, together with previous research, suggest that, on the day of delivery, the corpus luteum of the rat is in the very early stages of structural regression, with no changes at the morphological level, but with changes at the molecular level.